VALIDATION IMAGES
25 ug total protein per lane of various lysates (see on figure) probed with ATG7 monoclonal antibody, unconjugated (bsm-61206R) at 1:5000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
Paraformaldehyde-fixed, paraffin embedded Human Tonsil; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with ATG7 Monoclonal Antibody, Unconjugated (bsm-61206R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Spleen
; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with ATG7 Monoclonal Antibody, Unconjugated (bsm-61206R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
4% Paraformaldehyde-fixed HepG2 (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (ATG7) monoclonal Antibody, unconjugated (bsm-61206R) 1:100, 90 min at 37°C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37°C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.
ATG7 was immunoprecipitated from HeLa cell lysate with bsm-61206R at 1/100 dilution. Western blot was performed from the immunoprecipitate using protein A/G beads. HRP conjugated Goat Anti-Rabbit IgG was used as secondary antibody.
Lane 1: HeLa cell lysate (Input).
Lane 2: bsm-61206R IP in HeLa cell lysate.
Lane 3: native Rabbit IgG IP in HeLa cell lysate (negative control).